Haynes Lab:Notebook/CRISPR Editing/2015/03/24

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03/24/2015

Ran on gel and did PCR purification kit for unt and g025 treated luc14 and gal4+dox cekks, nested PCR with P173/215.

Surveyor hybridization protocol on 200ng in 15ul

Cell type Treatment Rep 260 280 260/280 ng/µL ul for 200ng ul hyb buff ul water
Luc14Unt10.0370.0191.8936.6045.461.508.04
Luc14Unt20.0250.0141.74525.1377.961.505.54
Luc14Unt30.0240.0131.79223.7048.441.505.06
Gal4-EED+doxUnt10.0270.0151.7926.7737.471.506.03
Gal4-EED+doxUnt20.0260.0141.87926.17.661.505.84
Gal4-EED+doxUnt30.0220.0121.87421.5549.281.504.22
Luc14g02510.0230.0121.89723.0038.691.504.81
Luc14g02520.0240.0131.76323.6538.461.505.04
Luc14g02530.0230.0141.64922.9358.721.504.78
Gal4-EED+doxg02510.0230.0131.7623.1118.651.504.85
Gal4-EED+doxg02520.0180.011.76517.93911.151.502.35
Gal4-EED+doxg02530.0260.0161.5926.087.671.505.83


Cut with 1ul Surveyor enzyme and 1ul enhancer. Ran on BA.


PCR purified

Cell type Treatment rep 260 280 260/280 ng/µL ul for 200ng ul of hyb buff ul water
Luc14Unt10.0370.0191.92236.6045.461.508.04
Luc14Unt20.030.0161.8430.1856.631.506.87
Luc14Unt30.0280.0151.84527.7257.211.506.29
Gal4-EED+doxUnt10.0330.0191.69932.5256.151.507.35
Gal4-EED+doxUnt20.0320.0191.67431.8886.271.507.23
Gal4-EED+doxUnt30.0280.0151.8327.8767.171.506.33
Luc14g04410.0280.0151.81828.1267.111.506.39
Luc14g04420.0310.0171.75330.5146.551.506.95
Luc14g04430.030.0171.73230.1996.621.506.88
Gal4-EED+doxg04410.0250.0141.76625.3517.891.505.61
Gal4-EED+doxg04420.0320.0171.8131.5226.341.507.16
Gal4-EED+doxg04430.0340.021.69633.5175.971.507.53
Luc14g04810.0260.0161.67526.4537.561.505.94
Luc14g04820.0180.0111.64818.22710.971.502.53
Luc14g04830.0230.0121.92122.7278.801.504.70
Gal4-EED+doxg04810.0270.0151.79526.6757.501.506.00
Gal4-EED+doxg04820.0220.0131.74222.3978.931.504.57
Gal4-EED+doxg04830.0280.0171.67327.6087.241.506.26

g023 and g046 gDNA extraction

Cell type Treatment rep 260 280 260/280 ng/µL ul for 200ng
Luc14g02310.1270.0641.983127.0481.57
Luc14g02320.1070.0551.948106.7521.87
Luc14g02330.0670.0341.96667.1962.98
Gal4-EED +doxg02310.0960.0472.05195.92.09
Gal4-EED +doxg02320.1030.052.067103.3471.94
Gal4-EED +doxg02330.0970.051.9597.2022.06
Luc14g04610.1510.0771.97151.1921.32
Luc14g04620.0980.0491.99298.0952.04
Luc14g04630.0930.0471.96993.42.14
Gal4-EED +doxg04610.1350.0691.959135.411.48
Gal4-EED +doxg04620.0860.0432.00286.3752.32
Gal4-EED +doxg04630.080.0421.92279.872.50


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